Investigation of nerve fibers in the skin by biopsy: technical aspects, indications, and contribution to diagnosis of small-fiber neuropathy.

Skin biopsy with investigation of small-diameter nerve fibers in human epidermis and dermis has been proven to be a useful method for confirming small-fiber neuropathy. In medical practice, small-fiber neuropathy is increasingly recognized as a leading cause of neuropathic pain. It is a prevalent complaint in medical offices, brought by patients often as a "painful burning sensation". The prevalence of neuropathic pain is high in small-fiber neuropathies of different etiologies, especially in the elderly; 7% of population in this age group present peripheral neuropathy. Pain and paresthesia are symptoms which might cause disability and impair quality of life of patients. The early detection of small-fiber neuropathy can contribute to reducing unhealthy lifestyles, associated to higher incidence of the disease.

Site-Specific Regulation of Sulfatase and Aromatase Pathways for Estrogen Production in Endometriosis.

Endometriosis is a highly prevalent gynecological disease characterized by lesions in different sites. Regulation of specific estrogen pathways may favor the formation of distinct microenvironments and the progression of endometriosis. However, no study has simultaneously evaluated the gene and protein regulation of the main estrogen-synthesizing enzymes in endometriosis. Thus, our goals were to study the relationship between gene and protein expression of aromatase (CYP19A1 or ARO), steroid sulfatase (STS), and hydroxysteroid 17-beta dehydrogenase (HSD17B1) in superficial (SUP), ovarian (OMA), and deep infiltrating (DIE) endometriotic lesion sites as well as in the eutopic endometrium of patients with (EE) and without (control) endometriosis in the same and large cohort of patients. The site-specific expression of these enzymes within different cells (glandular and stromal components) was also explored. The study included 108 patients surgically diagnosed with endometriosis who provided biopsies of EE and endometriotic lesions and 16 disease-free patients who collected normal endometrium tissue. Our results showed that CYP19A1 was detected in all endometriosis tissues and was in higher levels than in control. Unique patterns of the STS and HSD17B1 levels showed that they were most closely regulated in all tissues, with manifestation at greater levels in DIE compared to the other endometriotic lesion sites, OMA and SUP. Gene and protein expression of ARO, STS, and HSD17B1 occurred at different rates in endometriotic sites or EE. The distinctive levels of these estrogen-synthesizing enzymes in each endometriotic site support the hypothesis of a tissue microenvironment that can both influence and be influenced by the expression of different estrogenic pathways, locally affecting the availability of estrogen needed for maintenance and progression of endometriotic lesions.

Altered p16Ink4a, IL-1ß, and Lamin b1 Protein Expression Suggest Cellular Senescence in Deep Endometriotic Lesions.

Endometriosis causes immunological and cellular alterations. Endometriosis lesions have lower levels of lamin b1 than the endometrium. Moreover, high levels of pro-inflammatory markers are observed in the peritoneal fluid, follicular fluid, and serum in endometriosis lesions. Thus, we hypothesized that the accumulation of senescent cells in endometriosis tissues would facilitate endometriosis maintenance in an inflammatory microenvironment. To study senescent cell markers and the senescence-associated secretory phenotype (SASP) in endometriosis lesions, we conducted a cross-sectional study with 27 patients undergoing video laparoscopy for endometriosis resection and 19 patients without endometriosis. Endometriosis lesions were collected from patients with endometriosis, while eutopic endometrium was collected from patients both with and without endometriosis. Tissues were evaluated for senescence markers (p16Ink4a, lamin b1, and IL-1ß) and interleukin concentrations. The expression of p16Ink4a increased in lesions compared to that in eutopic endometrium from endometriosis patients in the secretory phase. In the proliferative phase, lesions exhibited lower lamin b1 expression but higher IL-4 expression than the eutopic endometrium. Further, IL-1ß levels were higher in the lesions than in the eutopic endometrium in both the secretory and proliferative phases. We believe that our findings may provide targets for better therapeutic interventions to alleviate the symptoms of endometriosis.

Investigation of nerve fibers in the skin by biopsy: technical aspects, indications, and contribution to diagnosis of small-fiber neuropathy

ABSTRACT Skin biopsy with investigation of small-diameter nerve fibers in human epidermis and dermis has been proven to be a useful method for confirming small-fiber neuropathy. In medical practice, small-fiber neuropathy is increasingly recognized as a leading cause of neuropathic pain. It is a prevalent complaint in medical offices, brought by patients often as a "painful burning sensation". The prevalence of neuropathic pain is high in small-fiber neuropathies of different etiologies, especially in the elderly; 7% of population in this age group present peripheral neuropathy. Pain and paresthesia are symptoms which might cause disability and impair quality of life of patients. The early detection of small-fiber neuropathy can contribute to reducing unhealthy lifestyles, associated to higher incidence of the disease.

Depleted lamin B1: a possible marker of the involvement of senescence in endometriosis?

PROPOSE: Endometriosis is a benign disease characterized by implantation and the growth of endometrial tissue outside the uterine cavity and it shares similarities with cancer. Lamin B1, p16 and p21 play a role on cell cycle regulation, development, cell repair and its activities are related to cancers. Considering the similarities between endometriosis and cancer, the aim of the present cross-sectional study is to detect p16, p21 and Lamin B1 in the ectopic endometrium of patients with endometriosis (n = 8) with eutopic (n = 8) and control endometrium (n = 8) and relate them to the maintenance and development of endometriosis. METHODS: Biopsies were obtained from both eutopic and ectopic, from deep infiltrating lesions, endometrium frozen and used for immunofluorescent (p16) or immunohistochemistry procedures (p16, p21, lamin B1). RESULTS: Detected higher lamin B1 in the eutopic endometrium when compared with ectopic endometrium, with no differences between endometriosis tissue with control endometrium. Similar presence of p16 in all groups of patients and no p21 detection was observed. CONCLUSION: We observed reduced detection of lamin B1 in the ectopic endometrium raising the possibility that the presence of senescent cells might be contributing to the maintenance and progression of endometriosis by apoptosis resistance and peritoneal stress inherent of the disease.

Cell-cycle analysis and apoptosis-associated proteins in cervical lesions of Brazilian women.

AIM: The aim of the present study was to detect the relative expressions of p53, p21(Waf1/Cip1), p27(Kip1) Bcl-2 and cleaved caspase-3 in cervical lesion samples from Brazilian women by immunohistochemistry. MATERIALS AND METHODS: A total of 230 cervical biopsies in paraffin-embedded blocks were studied: 43 were invasive squamous cell carcinomas (SCC), 52 carcinomas in situ/cervical intraepithelial neoplasias III (CIN III), 54 cervical intraepithelial neoplasias II (CIN II), 51 cervical intraepithelial neoplasias I (CIN I) and 30 non-neoplastic lesions (NN) with benign cellular changes. RESULTS: Significant differences were observed in the p53 expression between the different groups: NN and CIN I (p=0.010); NN and CIN II (p<0.00001); CIN II and CIN III (p=0.02); CIN II and CIS (p=0.0220); CIN II and CEC (p=0.010). Regarding p21(WAF1/Cip1), significant differences were observed between NN and CEC (p=0.001); CIN I and CEC (p=0.001); CIN II and CIN III (p=0,001); CIN II and CIS (p=0.0004) and CIN II and CEC (p<0.0001). For p27(Kip1), significant differences were observed between NN and CIN I (p<0.00001); NN and CIN II (p<0.00001); NN and CIS (p=0.038); CIN I and CIN III (p=0.001); CIN I and CIS (p=0.009); CIN I and CEC (p=0.0001); CIN II and CIN III (p=0.0003); CIN II and CIS (p=0.002); CIN II and CEC (p< 0.00001). Bcl-2 and caspase-3 did not show remarkable differences between groups. CONCLUSION: p53, p21(WAF1/CIP1), p27(KIP1) appear to be involved in the course of carcinogenesis. Rare expression of Bcl-2 and cleaved caspase-3 suggests that these proteins probably do not participate in cervical apoptosis.

HSP27 is commonly expressed in cervical intraepithelial lesions of Brazilian women.

Heat shock proteins are molecular chaperones that may be constitutively present in cells protecting them from various stresses, such as extreme temperature, anoxia or chemical agents. Cervical cancer is the second most prevalent malignancy of women. In this study, we analyzed the expression of Hsp27 by immunohistochemistry in cervical intraepithelial lesions of Brazilian women, along with samples from non neoplasic lesions (NN). Cervical intraepithelial neoplasia I (CIN I), II (CIN II) and III (CIN III)/in situ carcinoma and squamous cell carcinoma (SCC) were included. Immunostaining was observed in 30 (100%) samples of NN, 46 (92%) in CIN I, 50 (100%) in CIN II, 52 (98.11%) in CIN III/CIS, and 46 (98.11%) in SCC. In group NN Hsp27 immunostaining was heterogeneous, more intense in basal and parabasal layers of the epithelium and less or absent in the intermediate and superficial layer. The majority of the samples of CIS and SCC presented strong staining in allepithelial layers. Metaplasic cells, when present, were strongly stained. In this study, Hsp27 protein was found to be commonly expressed in cervical epithelial cells.

Correlation analysis of c-myc, p21(WAF/CIP1), p53, C-erbB-2 and COX-2 proteins in esophageal squamous cell carcinoma.

Esophageal cancer is one of the most frequently occurring malignancies and the seventh leading cause of cancer-related deaths in the world. Esophageal squamous cell carcinoma (ESCC) is the most common histological type of esophageal cancer worldwide. Our goal in this study was to detect c-myc, p21(WAF/CIP1), p53, C-erbB-2 and COX-2 immuno-expression in ESCC. Archival formalin-fixed, paraffin-embedded tissues of 18 cases of ESCC (13 biopsies and 5 surgical specimens) were studied, retrospectively, by immunohistochemistry. p53 protein was observed in 50% of the cases, while c-myc was found in 6 of 18 samples (33.33%). All samples (100%) were negative for p21(WAF/CIP1). C-erbB-2 oncoprotein and the COX-2 protein were detected in 5.5% (1/18) and 16.66% (3/18) of the cases, respectively. Taken together, our results suggest that c-myc, p53, C-erbB-2 and COX-2 proteins do not correlate with cancer stage or follow-up in ESCC as revealed by immunohistochemistry.

Matrix metalloproteinase-9 is consistently expressed in Hodgkin/Reed-Sternberg cells and has no impact on survival in patients with Epstein-Barr virus (EBV)-related and non-related Hodgkin lymphoma in Brazil.

Clinical and histological features of classical Hodgkin lymphoma (cHL) are primarily due to the effects of cytokines and enzymes produced by Hodgkin/Reed-Sternberg (HRS) cells and their surrounding inflammatory cells. In EBV-related cancers, the expression of viral latent membrane protein 1 correlates with an increased MMP9 expression. In this study, we evaluated the prognostic relevance of MMP9 expression and EBV status in HRS cells in patients with cHL in Brazil. We selected 97 patients with cHL for EBV and MMP9 detection. EBV was detected in 52.5%, and MMP9 expression positivity was found in 87.6%. Of all cases, there was no correlation between MMP9 expression and EBV status. Response to treatment and relapse rate was independent of MMP9 expression and EBV status. MMP9 positivity did not influence overall survival and event-free survival. The consistent and increased intensity of MMP9 expression in HRS cells make this enzyme a potential target for therapy.

Expression of p27, p21(WAF/Cip1), and p16(INK4a) in normal oral epithelium, oral squamous papilloma, and oral squamous cell carcinoma.

AIM: To characterize the immunohistochemical expression of p27, p21(WAF/Cip1) and p16(INK4a) in normal oral epithelium, oral squamous papilloma and oral squamous cell carcinoma. MATERIALS AND METHODS: Immunohistochemical staining for p27, p21(WAF/Cip1) and p16(INK4a) was evaluated in 32 samples of normal oral squamous epithelium, 30 samples of oral squamous papilloma, and 34 samples of oral squamous cell carcinoma. RESULTS: Statistically significant differences (p<0.05) were found in p27 expression when comparing ordinary mucosa and oral squamous papilloma with the oral squamous cell carcinoma samples. Regarding p21(WAF/Cip1) expression, no statistically significant differences (p>0.05) were noticed. In the same way, no significant statistically differences (p>0.05) were observed for p16(INK4a) among groups. CONCLUSION: Taken together, these findings indicate that p27 is closely involved in malignant transformation of oral mucosa cells, and may be a reliable biomarker for this purpose.

Impact of Epstein-Barr virus in the clinical evolution of patients with classical Hodgkin's lymphoma in Brazil.

INTRODUCTION: Classical Hodgkin's Lymphoma (cHL) has been frequently associated with Epstein-Barr virus (EBV), which can be found in a latent pattern in Reed-Sternberg (RS) cells. However, the impact of the presence of EBV in RS cells and its prognosis are still controversial. We analysed the presence of EBV in RS cells and its influence in the clinical evolution of patients with cHL treated in two public hospitals in the city of São Paulo, Brazil. MATERIALS AND METHODS: We selected 97 patients with cHL from 1994 to 2004. Patients were only included in this study if they had (1) >18 years, (2) negative HIV serology, (3) undergone similar chemotherapy protocols, (4) paraffin blocks available with enough material for systematic review and histological reclassification and for detection of EBV in RS cells by in situ hybridization and immunohistochemistry and (5) clinical, epidemiological and laboratorial parameters available after a thorough chart review. RESULTS: EBV was identified in 52.5% of the cases. Mixed cellularity (MC) subtype was more common in EBV-related tumours (25.5%) (p=0.005). There was no difference on age, gender, stage and the presence of B symptoms between the two groups. The presence of EBV did not influence event free survival (EFS) (p=0.38) or overall survival (OS) (p=0.80) with a median follow-up of 80 months. CONCLUSION: We demonstrate that the prevalence of EBV-related cHL in this Brazilian population is 52.5% and, that, the presence of EBV does not change the clinical evolution and OS of patients treated with similar chemotherapy protocols.

Analysis of GB virus C infection among HIV-HCV coinfected patients.

The aim of this study was to evaluate the effect of GB virus C on laboratory markers and histological parameters among HIV-seropositive patients coinfected with HCV. Lower degrees of hepatic lesions were observed in the triple-infected patients, in comparison with HIV-HCV coinfected patients who were negative for GBV-C RNA.

Estudo de p27, p21, p16 em epitélio escamoso normal, papiloma escamoso e carcinoma de células escamosas da cavidade oral / Comparative analysis of the immunohistochemistry expression of p27, p21WAF/Cip1, and p16INK4a in oral normal epithelium, squamous papilloma and squamous cell carcinoma

INTRODUÇÃO E OBJETIVO: O tipo de câncer oral mais frequente é o carcinoma de células escamosas, que corresponde a 95 por cento dos casos(9). O papiloma escamoso oral é uma neoplasia benigna normalmente associada à infecção pelo papilomavírus humano (HPV)(21). A análise da literatura mostra alterações nos genes reguladores do ciclo celular p27, p21WAF/Cip1 e p16INK4a, porém sem uma definição de seus papéis na carcinogênese oral. O objetivo foi caracterizar imuno-histoquimicamente p27, p21WAF/Cip1 e p16NK4a em epitélio escamoso normal, papilomas escamosos e carcinomas de células escamosas da cavidade oral. MÉTODOS: Imuno-histoquímica para p27, p21WAF/Cip1 e p16NK4a em 32 casos de epitélio escamoso normal, 30 casos de papiloma escamoso e 34 de carcinoma de células escamosas da cavidade oral. RESULTADOS: p27: 97,06 por cento dos casos de carcinoma de células escamosas apresentaram imunopositividade focal. O grupo papiloma escamoso apresentou 33,33 por cento e o grupo controle, 18,75 por cento. p21WAF/Cip1: 100 por cento de imunopositividade focal tanto no grupo controle como no grupo carcinoma de células escamosas, e 90 por cento no grupo papiloma escamoso. p16INK4a: 100 por cento de imunopositividade focal para os grupos controle e papiloma escamoso, e 94 por cento para o grupo carcinoma de células escamosas. CONCLUSÃO: Imuno-histoquimicamente demonstrou-se diferença significativa para p27 quando feita comparação dos grupos controle e papiloma escamoso com o grupo carcinoma de células escamosas. O p21WAF/Cip1 não demonstrou poder de diferenciar os grupos analisados. O p16INK4a apresentou imunopositividade difusa em uma minoria dos casos do grupo carcinoma de células escamosas. O grupo papiloma escamoso se comportou de maneira similar ao grupo controle em relação aos três marcadores.

INTRODUCTION: The most frequent type of oral cancer is the squamous cell carcinoma, which corresponds to 95 percent of the cases(9).The oral squamous papilloma is a benign neoplasia, commonly associated with infections caused by the human papilloma virus(21). The analysis of medical literature shows changes in cell cycle regulatory genes (p27, p21WAF/Cip1 and p16INK4a), but does not define their roles in oral carcinogenesis. Objective: Characterize the immuno-histochemical expression of p27, p21WAF/Cip1 and p16INK4a in oral normal squamous epithelium, oral squamous papilloma and oral squamous cell carcinoma. METHODS: Immuno-histochemical evaluation of p27, p21WAF/Cip1 and p16INK4a in 32 samples of oral normal squamous epithelium, 30 of oral squamous papilloma and 34 of oral squamous cell carcinoma. RESULTS: 97.06 percent of the oral squamous cell carcinoma group, 33.33 percent of the squamous papilloma group and 18.75 percent of the control group showed focal immunopositivity for p27. 100 percent of both control and oral squamous cell carcinoma groups and 90 percent of the oral squamous papilloma group showed focal immunopositivity for p21WAF/Cip1. 100 percent of both control and oral squamous papilloma groups and 94 percent of the oral squamous cell carcinoma group showed focal immunopositivity for p16INK4a. CONCLUSIONS: The study revealed a statistically significant difference for p27 expression when comparing the control and oral squamous papilloma groups with the oral squamous cell carcinoma group. p21WAF/Cip1 did not prove to be useful to differentiate the groups. p16INK4a showed diffuse immunopositivity in a minority of the oral squamous cell carcinoma cases. The oral squamous papilloma group behaved similarly to the control group as to the three markers.

Analysis of GB virus C infection among HIV-HCV coinfected patients / Análise da infecção pelo vírus GB-C em pacientes com coinfecção VIH-VHC

The aim of this study was to evaluate the effect of GB virus C on laboratory markers and histological parameters among HIV-seropositive patients coinfected with HCV. Lower degrees of hepatic lesions were observed in the triple-infected patients, in comparison with HIV-HCV coinfected patients who were negative for GBV-C RNA.

O objetivo do estudo foi avaliar o efeito da infecção pelo vírus GB-C em marcadores laboratoriais e parâmetros histológicos em pacientes HIV soropositivos coinfectados com VHC. Menor grau de lesão hepática foi observado nos pacientes com tripla infecção em comparação aos pacientes coinfectados com VIH-VHC negativos para GBV-C RNA.

Immunohistochemical expression of p16(INK4A) in normal uterine cervix, nonneoplastic epithelial lesions, and low-grade squamous intraepithelial lesions.

OBJECTIVES: In this study, the authors analyzed the immunoexpression of p16 in high-risk human papillomavirus DNA-negative normal and nonneoplastic cervical epithelia, in low-grade cervical intraepithelial neoplasia (CIN), high-grade CIN, and squamous cell carcinoma. MATERIALS AND METHODS: A retrospective study, in which 58 normal cervical hysterectomy samples, 56 nonneoplastic cervical biopsies, 88 CIN 1, 33 CIN 2, 32 CIN 3, and 47 invasive squamous cell carcinoma biopsies, were evaluated for p16 immunoexpression. Human papillomavirus tests were also performed. RESULTS: p16 immunohistochemistry seems to reveal possible different biological subgroups of lesions among morphologically similar mildly dysplastic cervical epithelia. CONCLUSION: Distribution patterns of p16 protein might be useful to predict different outcomes in CIN 1.

Método de hibridização in situ para detecção da infecção pelo papilomavírus humano (HPV) em regiões de vulva, vagina e colo uterino / Method of the in situ hybridization by detection of human papillomavirus (HPV) infection in vulva, Vagina and uterine cervix regions

O câncer cervical é. atualmente doença sexualmente transmissível mais comum entre mulheres jovens chegando a atingir 20 por cento a 46 por cento destas em vários países. O HPV corresponde ao principal fator etiológico da carcinogênese cervical. HPVs que induzem lesões que progridem para displasias e cânceres invasivos são chamados de vírus de alto risco (16,18), vírus de risco moderado (31,33) são também encontrados em lesões pré-malignas e vírus de baixo risco (6,11) estão mais comumente associados a lesões benigna do tipo condilomatosas. Para detecção da presença de DNA viral do HPV, foram estudadas 332 biópsia referentes ao ano de 1999 de uma amostra de 269 mulheres. Pela técnica de HIS, 113 (34,04 por cento) exames foram positivas para infecção pelo HPV, sendo 40 (24,39 por cento) das 164 biópsias de colo uterino, 46 (38,02 por cento das 121 biópsias de vulva e 27 (57,45 por cento) das 47 de vagina. O estudo da positividade das quatro sondas estudadas demonstrou significância entre os resultados obtidos nas regiões de colo uterino e vulva. Observamos maior número de positividade utilizando-se a sonda A/E nas três regiões. Nas lesões NlC obtivemos resultados positivos para pesquisa de infecção pelo HPV em 88,88 por cento (8/9) e 75 por cento (3/4) no casos de NIVA. Diferentes técnicas empregadas para a prevenção das neoplasias de colo uterino, vagins e vulva são importantes, mas somente a técnica de hibridização in situ pode correlacionar morfologicamente com maior especificidade a infecção pelo HPV e permitir análises retrospectivas em material arquivado. Os resultados demonstram que evidências morfológicas da infecção viral observadas pelo exame anatomopatológico são, na maioria, confirmadas pela técnica de HIS nessas localizações anatômicas.